Polymeric particles containing microorganisms

Abstract

The present invention relates to polymeric particles comprising a biodegradable polymer, and at least one microorganism in a total concentration of at least 108 CFU/g dry weight that is stable for at least 35 weeks at 30° C. and optionally additional carriers and additives as well as to methods for producing polymeric particles and use thereof.

The invention claimed is: 
     
       1. A polymeric composition, comprising polymeric particles which contain one or more microorganisms in one or more multicellular aggregates embedded in a self-produced extracellular biogenic matrix, wherein the self-produced extracellular biogenic matrix comprises naphthazarin, and wherein at least one microorganism has a total concentration of at least 10 8  CFU/g dry weight and storage at 30° C. for 14 weeks resulting in less than 1 log 10 loss CFU/g dry mass. 
     
     
       2. The polymeric composition of  claim 1 , wherein the composition comprises napthazarin in at least a 10 fold higher amount compared to a reference composition, wherein the reference composition is produced by culturing the microorganisms in a nutrient broth to a final concentration of 10 9  CFU/ml, encapsulating the microorganisms in alginate beads, incubating the beads for 24 to 48 hours in fresh nutrient broth, washing the alginate beads, and then either lyophilizing the alginate beads or placing the alginate beads in hermetically sealed containers. 
     
     
       3. The polymeric composition of  claim 1 , wherein the self-produced extracellular biogenic matrix additionally comprises one or more compound selected from the group consisting of homocysteine, NAD+/NADH, tryptophan, thiamine, erythrose 4-phosphate, phenylalanine, pyridoxine phosphate, and adenine. 
     
     
       4. The polymeric composition of  claim 3 , wherein the self-produced extracellular biogenic matrix additionally comprises one or more of homocysteine, NAD+/NADH, tryptophan, thiamine, erythrose 4-phosphate, phenylalanine, pyridoxine phosphate, or adenine in at least a 1 fold higher amount compared to a reference composition, wherein the reference composition is produced by culturing the microorganisms in a nutrient broth to a final concentration of 10 9  CFU/ml, encapsulating the microorganisms in alginate beads, incubating the beads for 24 to 48 hours in fresh nutrient broth, washing the alginate beads, and then either lyophilizing the alginate beads or placing the alginate beads in hermetically sealed containers. 
     
     
       5. The polymeric composition of  claim 1 , wherein the polymeric particles are wet and at least 1 of the one or more multicellular aggregates is between 14 μm and 43 μm in diameter. 
     
     
       6. The polymeric composition of  claim 1 , wherein the polymeric particles comprise a biodegradable polymer, wherein the biodegradable polymer is selected from the group consisting of albumin, collagen, gelatin, fibrinogen, casein, fibrin, hemoglobin, transferrin, chitin, chitosan, hyaluronic acid, heparin, chondroitin, keratin sulfate, alginate, starch, dextrin, dextran, polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, polyhydroxybutyric acid, polycaprolactone, polyanhydride, agarose, agar, chitosan, and polyalkylcyanoacrylate. 
     
     
       7. The polymeric composition of  claim 1 , wherein at least one of the polymeric particles has a diameter of less than 200 μm. 
     
     
       8. The polymeric composition of  claim 1 , wherein the one or more encapsulated microorganisms are bacterial or fungal cells. 
     
     
       9. The polymeric composition of  claim 1 , wherein the one or more encapsulated microorganisms comprise at least two microorganisms of distinct genetic origins. 
     
     
       10. The polymeric composition of  claim 9 , wherein the at least two microorganisms of distinct genetic origins are selected from at least two of the group archaea, protozoa, bacteria, fungi and algae. 
     
     
       11. The polymeric composition of  claim 1 , wherein the viability of at least one microorganism is maintained for a storage period of at least 35 weeks at 30° C.